Antigens, CD20 ; metabolism ; CD79 Antigens ; metabolism ; Heart Atria ; Heart Neoplasms ; diagnostic imaging ; metabolism ; pathology ; surgery ; Humans ; Immunohistochemistry ; Lymphoma, Large B-Cell, Diffuse ; diagnostic imaging ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Tomography, X-Ray Computed

OBJECTIVETo investigate the effect of RNA interference (RNAi)-mediated silencing of the SREBP2 on inflammatory cytokine-induced cholesterol accumulation in HepG2 cells.

METHODSShort-hairpin (sh)RNA targeting SREBP2 or negative control (NC) shRNA were transfected into HepG2 cells by a liposomal method. G418-selective culturing was used to obtain the SREBP2 shRNA HepG2 and NC shRNA HepG2 cell lines. The two cell lines were cultured in serum-free medium and left untreated (control) or treated with TNF-a (20 ng/ml), low-density lipoprotein (LDL) loading (100 mug/ml), or a combination LDL plus TNF-a treatment. Lipid accumulation was evaluated by oil red O (ORO) staining. Intracellular cholesterol level was measured by enzymatic assay. The mRNA and protein levels of SREBP2 and its downstream target genes, LDL receptor (LDLr), and HMGCoA reductase, were measured by real-time PCR and Western blotting, respectively.

RESULTSSREBP2 shRNA HepG2 and NC shRNA HepG2 stable cell lines were successfully established. ORO staining and cholesterol quantitative analysis showed that LDL loading significantly increased intracellular cholesterol and that expression of SREBP2 further exacerbated the inflammatory cytokine-induced lipid accumulation, as seen in NC shRNA HepG2 cells. LDL loading of NC shRNA HepG2 decreased the gene and protein expressions of SREBP2, LDLr, and HMGCoA reductase, but the suppressive effect was overridden by inflammatory cytokine. SREBP2 shRNA HepG2 cells showed lower levels of cholesterol accumulation under LDL loading and inflammatory stress conditions. Moreover, the mRNA and protein levels of SREBP2, LDLr, and HMGCoA reductase were much lower than in NC shRNA HepG2 cells under the same conditions.

CONCLUSIONInflammatory cytokine exacerbated cholesterol accumulation in HepG2 via disrupting SREBP2. RNAi-mediated inhibition of SREBP2 expression significantly ameliorated the cholesterol accumulation induced by inflammatory cytokine.

Cholesterol ; metabolism ; Hep G2 Cells ; Humans ; Inflammation ; RNA Interference ; RNA, Small Interfering ; Sterol Regulatory Element Binding Protein 2 ; genetics ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo observe the effect of Aike Mixture (AKM) on prostatic inflammatory infiltration in patients with chronic prostatitis type III A (III A-CP/CPPS) and evaluate its anti-inflammatory action. METHODS METHODS: A total of 60 patients with III A-CP/CPPS suitable to operation and differentiated as Chinese medicine: Gan qi stagnancy syndrome type were selected. They were assigned with the random number table to two groups equally. Before operation, the patients in the treated group were administered with Proscar combined with AKM, but those in the control group treated with Proscar only. Suprapubic transvesical prostatectomy was performed two weeks later, and prostatic pathological examination was conducted.

RESULTSGrading of: inflammatory cell infiltration showed that the mean grade in the treated group was 0.78 ± 0.90 grades, which was significantly lower than that in the control group 1.68 ± 0.87 grades (P<0.05). However, the two groups were not different in the grades of fibroblast proliferation (1.50 ± 0.70 grades vs 1.62 ± 0.87 grades, P>0.05).

CONCLUSIONAKM could suppress the inflammatory cell infiltration, be an effective and safe remedy for the treatment of IIIA-CP/CPPS of Gan-qi stagnancy syndrome type, and worthy for spreading in clinical use.

Aged ; Chronic Disease ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Fibroblasts ; pathology ; Humans ; Hyperplasia ; Inflammation ; drug therapy ; pathology ; Male ; Middle Aged ; Prostatitis ; classification ; drug therapy ; pathology

Objective:To evaluate the gene mutation profile and prognostic significance of adult cytogenetically normal acute myeloid leukemia (CN-AML) with CEBPA-bZIP mutation. Methods:Targeted sequencing was implemented on the diagnostic bone marrow DNA samples of 141 adult CN-AML subjects with CEBPA-bZIP mutation. The nomogram model for leukemia-free survival (LFS) rate was generated by combining genetic abnormalities and clinical data. Risk stratification was conducted based on prognostic variables and the effect of risk-adjusted consolidation therapy was investigated by Kaplan-Meier method. Results:Four variables were finally included in our nomogram model after multivariate Cox analysis,and an equation for risk score calculation was obtained,risk score=1.3002×white blood cell (WBC) (≥18.77×109/L)+1.4065×CSF3R mutation positive+2.6489×KMT2A mutation positive+1.0128×DNA methylation-related genes mutation positive. According to the nomogram model,patients were further divided into low-risk group (score=0,n=46) and high-risk group (score>0,n=95). Prognostic analysis showed that the 5-year LFS rate,5-year overall survival (OS) rate,and 5-year cumulative incidence of relapse (CIR) of patients who received allogeneic hematopoietic stem cell transplantation (allo-HSCT) in the high-risk group were 93.5％,97.1％,and 3.5％,while those in patients who received maintenance chemotherapy were 32.9％,70.5％,and 63.4％,respectively. The differences were statistically significant (all P<0.05). Allo-HSCT could significantly improve the prognosis of patients in high-risk group. However,no corresponding benefit was observed in the low-risk group. Conclusion:Adult CN-AML with CEBPA-bZIP mutation has a complex co-mutation pattern. The nomogram model based on mutations of CFS3R,KMT2A and DNA methylation-related genes together with WBC count can further divide this subset of patients into a relatively low-risk group and a relatively high-risk group. For individuals in the high-risk group,allo-HSCT is proposed as post-remission therapy. The above data will benefit the prognosis estimation and treatment decision for adult CN-AML with CEBPA-bZIP mutation.

AIMTo study the therapeutic effects of oxyphenamone, a novel inodilator, on myocardial ischemia.

METHODSThe cardiac hemodynamic variables in cats with acute myocardial infarction induced by occlusion of the left anterior descending coronary artery (LAD) were recorded with a physiological polygraph and electromagnetic flowmeter. A model of myocardial necrosis induced by subcutaneous injection of isoproterenol was used for evaluating the effects of drugs on myocardial enzymes and morphological change.

RESULTSIntravenous injection of oxyphenamone (2 - 8 mg x kg(-1)) dose-dependently decreased heart rate, mean arterial pressure, vascular resistance and the parameters of myocardial oxygen consumption (tension time index, TTI) in cats with myocardial infarction. It increased myocardial contractile force and cardiac output transiently but showed no influence on the left ventricular pressure and cardiac work. The changes of myocardial morphology, creatine phosphate kinase (CPK), malodialdehyde (MDA) and serum glutamic-oxaloacetic transaminase (GOT) induced by isoproterenol in rats were diminished by intraperitoneal injection of oxyphenamone (4 - 8 mg x kg(-1)).

CONCLUSIONBy the examination of the cardiac hemodynamics, myocardial enzymes and morphology, it showed that the myocardial damage induced by ischemia or beta-agonist can be antagonized markedly by oxyphenamone, indicating that oxyphenamone may be beneficial for the treatment of myocardial infarction.

Animals ; Blood Pressure ; drug effects ; Cardiac Output ; drug effects ; Cardiotonic Agents ; pharmacology ; Cats ; Heart ; physiopathology ; Heart Rate ; drug effects ; Male ; Myocardial Contraction ; drug effects ; Myocardial Infarction ; pathology ; physiopathology ; Myocardium ; metabolism ; pathology ; Organic Chemicals ; pharmacology ; Rats ; Rats, Wistar ; Vasodilator Agents ; pharmacology

This study was purposed to investigate the effect of adenovirus-mediated transfer of PDCD5 gene on apoptosis of K562 cells induced by etoposide. Recombinant adenovirus PDCD5 (Ad-PDCD5), control vectors Ad-null and Ad-eGFP were constructed by AdMax vector system respectively. After K562 cells were transfected by Ad-PDCD5, Ad-null or Ad-eGFP with different multiplicity of infection (MOI), the expression level of the PDCD5 gene was examined by RQ-RT-PCR assay. The effects of etoposide in combination with Ad-PDCD5 on the proliferation and apoptosis of K562 cells were measured by using MTT assay and flow cytometry with Annexin-V-FITC/PI dual labeling technique, respectively. The results showed that the transfection efficiencies of Ad-eGFP in K562, Jurkat and CEM cells ranged from 60% to 86%. Expression level of PDCD5 gene in K562 cells was evidently increased following transfection with Ad-PDCD5. The Ad-PDCD5 synergistically enhanced the apoptotic percentage of K562 cells induced by VP-16, as compared with that of Ad-null + VP16 and VP-16 alone respectively. It is concluded that Ad-PDCD5 may be a potential agent for enhancing the chemotherapy effect.
Adenoviridae ; genetics ; metabolism ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; genetics ; metabolism ; Etoposide ; pharmacology ; Humans ; K562 Cells ; Neoplasm Proteins ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Transfection

OBJECTIVETo evaluate levels of common specific fusion transcripts M-bcr-abl, m-bcr-abl, TEL-AML1, AML1-ETO, PML-RAR alpha, CBF beta-MYH11 in untreated leukemia patients.

METHODSSpecific fusion transcript levels were detected by TaqMan-based real-time quantitative RT-PCR technique in a total of 208 samples, including 195 bone marrow samples from 50 M-bcr-abl(+) chronic phase-chronic myeloid leukemia (CML-CP), 10 M-bcr-abl(+) acute lymphoblastic leukemia (ALL), 19 m-bcr-abl(+) ALL, 11 TEL-AML1(+) ALL, 30 AML1-ETO(+) acute myeloid leukemia (AML), 58 PML-RAR alpha(+) acute promyelocytic leukemia (APL) and 17 CBF beta-MYH11(+) AML patients and 13 peripheral blood samples from 13 M-bcr-abl(+) CML-CP patients. abl was chosen as internal control gene. Fusion transcript level was calculated as fusion transcript copies/abl transcript copies in percentage.

RESULTSBone marrow and peripheral blood samples of CML-CP patients had similar M-bcr-abl fusion transcript levels (median 30% vs 35%, P > 0.05). M- and m-bcr-abl (median 64% vs 54%) levels were similar in ALL patients (P > 0.05), M-bcr-abl level was significantly higher in ALL than CML-CP patients(P < 0.001). Median TEL-AML1 level was 228% in ALL patients. Among AML patients, AML1-ETO level was significantly higher than CBF beta-MYH11 and PML-RAR alpha levels (median 388% vs 145%, 388% vs 47%, all P < 0.001), CBF beta-MYH11 level was significantly higher than PML-RAR alpha level (P < 0.001). Fusion transcript levels of L-, V- and S-type PML-RAR alpha were 45%, 44% and 55%, respectively. L-type was significantly lower than S-type (P = 0.04).

CONCLUSIONSFusion transcript levels in untreated leukemia patients were different and patient-to-patient variations did exist. Detection of fusion transcript levels in untreated leukemia patients not only provides baseline for minimal residual disease monitoring and treatment evaluation but also enable the comparison in inter-laboratory data.

Adolescent ; Adult ; Bone Marrow Cells ; metabolism ; Child ; Child, Preschool ; Core Binding Factor Alpha 2 Subunit ; genetics ; Female ; Fusion Proteins, bcr-abl ; genetics ; Humans ; Leukemia ; genetics ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; genetics ; RUNX1 Translocation Partner 1 Protein ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Transcription, Genetic

This study was aimed to investigate abca5, mdr-1, kdr, dapk and irf-1 expressions in leukemia stem/progenitor cells (LSC) from CD7 positive acute myeloid leukemia, the expression of these 5 genes in mononuclear cells (MNC) from 15 normal bone marrow (NBM) and 16 AML patients bone marrow (AML BM) specimen were detected by real-time quantitative PCR (RQ-PCR). CD34(+)CD38(+) progenitor cells and CD34(+)CD38(-)Lin(-) stem cells were sorted by flow cytometry (FCM) from the MNCs of 10 NBM and 21 AML BM specimen. These 5 gene expressions in the sorted cells were detected by small amount cell RQ-PCR. The results showed that these 5 genes above mentioned all expressed in NBM-MNC, in which the expression levels of irf-1 and dapk were highest with the relative expression levels 4.08 and 3.86, the expression levels of abca 5 and mdr-1 were in the middle with the relative expression 0.49 and 0.84 respectively, the kdr expression was lowest with the relative expression level 0.02. In CD34(+)CD38(+) progenitor cells, the expression level of kdr increased dramatically (p < 0.05) while irf-1 and dapk dramatically decreased (p < 0.05). There was no obvious change of expression in the rest 2 genes. In CD34(+)CD38(-) stem cells the expression level of these 5 genes all increased nearly 2 times as much as that in CD34(+)CD38(+) progenitor cells, but kdr increased 3 times as much, and the increase of kdr and irf-1 expressions was of statistical significance (p < 0.05). Compared with the NBM, expression levels of 5 genes in AML-MNC decreased, and out of them abca 5, mdr-1, kdr and dapk were decreased most remarkably (p < 0.05). Comparison between AML CD34(+)CD38(+) cells and AML MNC showed that the expression level of irf-1 and dapk were decreased dramatically (p < 0.05) while the rest 3 genes increased their expression with statistical significance (p < 0.05). The expression levels of these 5 genes were higher in CD34(+)CD38(-) cells than those in CD34(+)CD38(+) stem cells, and the increase of kdr and irf-1 expressions showed statistical difference (p < 0.05). These 5 genes expression levels were all higher than those in CD34(+)CD38(+) cells whether in AML CD34(+)CD38(-)CD7(+) cells or CD34(+)CD38(-)CD7(-) cells. The increase of kdr expression in CD34(+)CD38(-)CD7(+) cells as well as kdr and irf-1 expressions in CD34(+)CD38(-)CD7(-) cells were all of statistical significance (p < 0.05). In conclusion the expression level of kdr in NBM was highest in stem cells while dapk and irf-1 were highest in differentiated cells. The expression levels of these 5 genes in CD34(+)CD38(-)Lin(-) stem cells were higher than those in CD34(+)CD38(+) progenitor cells. The gene expressions in AML CD34(+)CD38(-)CD7(+) cells and CD34(+)CD38(-)CD7(-) cells are in accordance with the characteristics of stem cells.
Adolescent ; Adult ; Aged ; Antigens, CD7 ; immunology ; Bone Marrow Cells ; chemistry ; immunology ; Case-Control Studies ; Female ; Flow Cytometry ; Gene Expression Regulation, Leukemic ; Hematopoietic Stem Cells ; chemistry ; immunology ; Humans ; Leukemia, Myeloid, Acute ; genetics ; immunology ; Male ; Middle Aged ; Stem Cells ; chemistry ; immunology ; Young Adult

This study was aimed to summarize and analyze the morphology, immunophenotype, cytogenetics, molecular biology (MICM), tyrosine kinase (TK) gene mutations and clinical features of acute myeloid leukemia (AML) with complex variant of t(8;21). A retrospective study was performed for 20 AML patients with complex variant of t(8;21) in our hospital from January 1994 to April 2012, including analysis of clinical feature, immunophenotype, chromosome karyotype, treatment regimen, as well as the overall survival (OS) and relapse-free survival (RFS). Mutations of C-KIT, FLT3-ITD, FLT3-TKD and JAK2V617F were detected by genomic DNA PCR and the sequencing was per-formed in 13 AML patients with complex variant of t(8;21). The results showed that (1) the incidence of 20 AML patients with complex variant of t(8; 21) was 2.4% of total t(8; 21) AML patients. In 20 AML patients with complex variant of t(8;21), 1 case was M1, 17 cases were M2, 2 cases were M4; 10 cases were myeloid phenotype and the other 3 were myeloid plus lymphoid phenotype. There were 16 kinds of cytogenetics additional involvement of chromosomal breakpoints: lp22, 1p32, 2q35, 2q14, 3p25, 5q13, 6p22, 7q21, llq11, 1lq13, 12q14, 12q24, 12p12, 14q32, 15p13, 20q12. (2) C-KIT aberrations were detected in 30.8% cases, all mutated in exon 17 (mutkit 17), only 1 case had JAK2V617F mutation. The result of FLT3 mutation screenings in AML patients with complex variant of t(8; 21) was negative. Of 5 patients with gene mutations, 1 patient (20%) achieved complete remission (CR), the median RFS and median OS time were 6.5 months and 8.9 months respectively. Of the 8 patients without gene mutations, 6 patietns (75%) achieved CR; the median RFS and median OS time were 26.6 months and 27.7 months respectively. It is concluded that the AML patients with complex variant of t(8;21) shows typical features of t(8;21) AML, but the existence of the tyrosine kinase-related gene mutation has important implications on remission rate and long-term survival of patients treated by induction chemotherapy.
Adolescent ; Adult ; Aged ; Chromosomes, Human, Pair 21 ; Chromosomes, Human, Pair 8 ; DNA Mutational Analysis ; Female ; Humans ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Protein-Tyrosine Kinases ; genetics ; Retrospective Studies ; Translocation, Genetic ; Young Adult

The aim of this study was to investigate the gene expression of programmed cell death 5 (pdcd5) in plasma and bone marrow cells from patients with multiple myeloma (MM). Enzyme liked immunosorbent assay (ELISA) and real-time quantitative reverse transcription polymerase chain reaction (RQ-RT-PCR) were used to examine pdcd5 gene expression in plasma and marrow cells in 45 MM patients and 20 normal controls. The results showed that serum levels of PDCD5 protein in 45 MM patients were lower significantly compared with the normal controls and 20 responsive patients after chemotherapy, their plasma levels were (16.91 +/- 0.28) ng/ml, (19.11 +/- 0.29) ng/ml and (17.94 +/- 0.154) ng/ml respectively (p < 0.05). The pdcd5 gene expression levels detected by RQ-RT-PCR in 45 MM patients were lower significantly compared with the normal controls, their pdcd5 gene expression levels were 0.64 +/- 0.47 and 1.28 +/- 1.21 respectively (p < 0.05). It is concluded that the PDCD5 protein expression levels are low in patients with MM. These findings suggest that abnormal expression of pdcd5 may be involved in the pathogenesis of MM.
Adult ; Aged ; Apoptosis Regulatory Proteins ; genetics ; Bone Marrow Cells ; pathology ; Case-Control Studies ; Female ; Gene Expression ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; pathology ; Neoplasm Proteins ; genetics ; Young Adult